The Cell Biology and Imaging Core (Core B) which will be used by all of the Projects will be involved in[unreadable] three primary functions:[unreadable] 1) preparation of neonatal rat ventricular myocytes (NRVMs)[unreadable] 2) preparation of adult mouse ventricular myocytes (AMVMs)[unreadable] 3) live cell imaging studies[unreadable] 4) distribution and analysis of results through use of the Open Microscopy Environment (OME) in[unreadable] coordination with The Scripps Research Institute (TSRI)[unreadable] The Cell Biology and Imaging Core (Core B) which will be used by all of the Projects will be involved in three[unreadable] primary functions: 1) preparation of neonatal rat ventricular myocytes (NRVMs), 2) preparation of[unreadable] adult mouse ventricular myocytes (AMVMs), and 3) live cell imaging studies. Consistent preparation of[unreadable] cardiomyocytes is critical to the ability to integrate findings across the Projects in the Program. Accordingly,[unreadable] while many of the individual laboratories have experience or capability in this regard, Core B will serve as a[unreadable] repository to insure cell availability and cross integration. Preparation of stable neonatal rat ventricular[unreadable] myocytes is relatively simple, but experimental observations are highly dependent on the methodology in[unreadable] particular the separation of cardiomyocytes from fibroblasts, plating density and choice of culture medium.[unreadable] With regard to adult mouse ventricular cardiomyocytes the mechanics and success of the preparation,[unreadable] and in particular the yield and long term viability of these cells can require skill and consistency. A dedicated[unreadable] technician and facility is therefore prerequisite to providing cells from TG and KO mice as required by[unreadable] virtually every project. Core C will also facilitate in the development of methods for expressing genes through[unreadable] adenoviral infection and expressing proteins through preparation and delivery of TAT fusion proteins. Live[unreadable] cell imaging will also be performed by Core B. In particular, technical advice and assistance with[unreadable] experiments using the FRET based Akt activity probe BKAR will be provided, as will analysis of[unreadable] mitochondrial membrane potential (TMRE) and integrity (RIRR, Calcein). Support for analysis of autophagy[unreadable] and mitophagy using TAT or adenoviral LC3 or cardiomyocytes from LC3 transgenics will also be provided.[unreadable] Inverted fluorescence and confocal microscopes equipped for these analyses are available at both UCSD[unreadable] and TSRI. The Open Microscopy Environment (OME) operated through TSRI provides a unique opportunity[unreadable] for data sharing and analysis.